ClearLLab LS Lymphoid Screen Casebook
Introduction
This casebook has been designed to assist in the analysis of flow cytometric immunophenotyping data generated using Beckman Coulter’s ClearLLab LS Lymphoid Screen reagent on the Beckman Coulter Navios and Navios EX flow cytometers. Cases with characteristic findings typical of various lymphoid and myeloid neoplasms are included, as are cases from patients with clinical and/or laboratory findings that suggest an underlying neoplastic process, but in which no immunophenotypic abnormality is identified. Specimen types include peripheral blood, bone marrow, and lymph nodes.
Each case includes a clinical vignette that describes the patient demographics and clinical history, case-specific listmode data files for reanalysis by the user of this casebook, ClearLLab LS-specific analysis protocols to be used with the listmode data, and a report showing the analysis with provided protocols. Each report includes analysis notes that highlight the immunophenotypic findings as well as potential pitfalls.
NOTE: Casebook examples are provided for illustrative purposes only, and not all categories of hematolymphoid neoplasms may be represented, nor are all possible immunophenotypic variants described or demonstrated.
Background
Flow cytometric immunophenotyping evaluates the presence and absence of specific antigens for each individual cell present in the specimen. When taken together, these results generate an immunophenotypic profile for each cell which is either consistent with an expected population (i.e. normal) or inconsistent with an expected population (i.e. aberrant) in that sample type. When evaluating samples from patients with suspected hematolymphoid malignancies, several steps are involved1:
- Assessment of all cell populations in the sample.
- Assignment of each cell population to either “normal” or “aberrant”.
- Detailed characterization of the aberrant population according to the presence or absence of antigens as well as increased or decreased intensity of staining by fluorochrome-labeled antibodies.
- Interpretation of the aberrant immunophenotype, incorporating where available additional information such as clinical history, histology, cytology, immunohistochemistry, and genotyping studies such as in situ hybridization, karyotyping, and molecular diagnostics.
Consensus Recommendations for Immunophenotyping
Consensus recommendations for flow cytometric immunophenotyping of samples from patients with known or suspected hematolymphoid malignancies have emerged over the last two decades, and several guidelines have been published in the scientific literature. Flow cytometric immunophenotyping has been included in the WHO classification of Tumors of Haematopoetic and Lymphoid Tissues since 20082. Medical indications and flow cytometry assay validation including pre-analytic, analytic, and post-analytic details of testing are addressed in the 2006 Bethesda International Consensus Conference recommendations3,4,5 and the ISCH/ICCS practice guidelines for cell-based fluorescence assays6,7,8.
ClearLLab LS Lymphoid Screen Reagent Intended Use
ClearLLab LS (Lymphoid Screen) reagent is intended for in vitro diagnostic use as a screening panel for identification of various hematolymphoid cell populations by immunophenotyping on Navios and Navios EX flow cytometers. This reagent is used as an aid in the differential diagnosis of patients with signs and/or symptoms of hematolymphoid malignancies. The reagent can be used with peripheral whole blood (collected in EDTA, ACD or Heparin), bone marrow (collected in EDTA, ACD, or Heparin) and lymph node specimens for immunophenotyping. The results should be interpreted along with additional clinical and laboratory findings. These reagents provide qualitative results for T, B and NK lineages.
ClearLLab LS Lymphoid Screen Reagent (part number B74073)
1Pacific Blue. 2APC-Alexa Fluor 700. 3APC-Alexa Fluor 750.
The above reagent is provided in a standardized format to be used along with reagents for sample preparation and cytometer set-up, along with software for data acquisition and analysis. ClearLLab LS reagent meets recommendations for standardization as outlined by the Bethesda guidelines2. Additional information regarding ClearLLab LS Lymphoid Screen Reagent is available here.
Cases
The listmode data presented in this case book were generated following the procedure detailed within the ClearLLab LS Lymphoid Screen reagent Instructions For Use (IFU) available at beckman.com.
Representative cases were selected from clinical trial data and were reviewed, anotated, and interpreted by Hematopathologist, Jeannine T. Holden MD MBA, and former Director of Scientific Affairs for Beckman Coulter Inc. View the protocol file and explore the listmode data files linked within the cases below
References
- Flow Cytometric Immunophenotyping for Hematologic Neoplasms. F.E. Craig, K.A. Foon. Blood. 2008; 111; 3941-3967.
- Swerdlow SH, Campo E, Harris NL, Jaffe EA, Pileri SA, Stain H, Thiele J, & Vardiman JW (eds) (2008) WHO Classification of Tumors of Haematopoietic and Lymphoid Tissues. IARC Press: Lyon
- 2006 Bethesda International Consensus recommendations on the immunophenotypic analysis of hematolymphoid neoplasia by flow cytometry: optimal reagents and reporting for the flow cytometric diagnosis of hematopoietic neoplasia. Wood BL, Arroz M, Barnett D, DiGiuseppe J, Greig B, Kussick SJ, Oldaker T, Shenkin M, Stone E, Wallace P. Cytometry B Clin Cytom. 2007;72 Suppl 1:S14-22
- 2006 Bethesda International Consensus recommendations on the flow cytometric immunophenotypic analysis of hematolymphoid neoplasia: medical indications. Davis BH, Holden JT, Bene MC, Borowitz MJ, Braylan RC, Cornfield D, Gorczyca W, Lee R, Maiese R, Orfao A, Wells D, Wood BL, Stetler-Stevenson M. Cytometry B Clin Cytom. 2007;72 Suppl 1:S5-13
- 2006 Bethesda International Consensus Conference on Flow Cytometric Immunophenotyping of Hematolymphoid Neoplasia. Stetler-Stevenson M, Davis B, Wood B, Braylan R. Cytometry B Clin Cytom. 2007;72 Suppl 1:S3
- Validation of cell-based fluorescence assays: practice guidelines from the ICSH and ICCS - part III - analytical issues. Tanqri S, Vall H, Kaplan D, Hoffman B, Purvis N, Porwit A, Hunsberger B, Shankey TV; ICSH/ICCS Working Group. Cytometry B Clin Cytom. 2013 Sep-Oct;84(5):291-308. doi: 10.1002/cyto.b.21106
- Validation of cell-based fluorescence assays: practice guidelines from the ICSH and ICCS - part IV - postanalytic considerations. Barnett D, Louzao R, Gambell P, De J, Oldaker T, Hanson CA; ICSH/ICCS Working Group. Cytometry B Clin Cytom. 2013 Sep-Oct;84(5):309-14. doi: 10.1002/cyto.b.21107
- Validation of cell-based fluorescence assays: practice guidelines from the ICSH and ICCS - part V - assay performance criteria. Wood B, Jevremovic D, Béné MC, Yan M, Jacobs P, Litwin V; ICSH/ICCS Working Group. Cytometry B Clin Cytom. 2013 Sep-Oct;84(5):315-23. doi: 10.1002/cyto.b.2110
